What do you use enzymes for, in antibody identification?
to help separate the specificities and allow for identification
What antibodies are neutrallized with saliva, plasma, or serum?
Lewis Antibodies
How is adsorption used in antibody identification?
add a known antigen to the serum, centrifuge and remove antigen/antibody complex, test adsorbed serum for antibodies
What does alloadsorption remove?
autoantibodies and alloantibodies
What does autoadsorption do?
removes autoantibodies (cold and warm)
What is the procedure for autoadsorption?
- Incubate cells w/ patient plasma/serum for 1 hr
1a. Cold autoantibodies at 4C
1b.. Warm autoantibodies at 37C - If agglutination is present, incubate plasma/serum with new aliquot of autologous RBCs
What is DAT used to detect…
in vivo sensitization of RBCs
What is the procedure for DAT antibody identification?
- RBCs are washed 3x
- AHG is added
2a. Polyspecific AHG, then monospecific after
What is done after IgG are detected by DAT?
Eluate (separate Ab from RBC, then use eluate panel to identify the Ab)
What is the Eluate procedure?
- Heat RBCs at 45C
- Acid elution w/ glycine acid solution at 3 pH
- Organic acids
- Wash
- Test Eluate and Last wash supernatant (control)
Titer level is the _______ of the greatest dilution
reciprical
Why is initial titration frozen?
to test against new specimens as a variability control
(see if technician is doing shit different [ wants similar techniques everytime])
What is titration used for?
monitoring patients for transplants and potential HDFN
What do you phenotypically match for with sickle cell and thalassemic patients?
Rh and K antigens
